Spirocycles as Inhibitors of 11-beta Hydroxyl Steroid Dehydrogenase Type 1

ABSTRACT

The present invention relates to certain spirocyclic compounds that are inhibitors of 11-β hydroxyl steroid dehydrogenase type 1 (11βHSD1), compositions containing the same, and methods of using the same for the treatment of diabetes, obesity and other diseases.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.14/680,745, filed Apr. 7, 2015, which is a continuation of U.S. patentapplication Ser. No. 13/619,160, filed Sep. 14, 2012, now U.S. Pat. No.9,006,260, which is a continuation of Ser. No. 12/143,427, filed Jun.20, 2008, now U.S. Pat. No. 8,278,318, which claims the benefit of U.S.Ser. No. 60/945,487, filed Jun. 21, 2007. The entire disclosure of eachof the related applications is incorporated herein by reference in itsentirety.

FIELD OF THE INVENTION

The present invention relates to certain spirocyclic compounds that areinhibitors of 11-β hydroxyl steroid dehydrogenase type 1 (11βHSD1),compositions containing the same, and methods of using the same for thetreatment of diabetes, obesity and other diseases.

BACKGROUND OF THE INVENTION

The importance of the hypothalamic-pituitary-adrenal (HPA) axis incontrolling glucocorticoid excursions is evident from the fact thatdisruption of homeostasis in the HPA axis by either excess or deficientsecretion or action results in Cushing's syndrome or Addison's disease,respectively (Miller and Chrousos (2001) Endocrinology and Metabolism,eds. Felig and Frohman (McGraw-Hill, New York), 4^(th) Ed.: 387-524).Patients with Cushing's syndrome (a rare disease characterized bysystemic glucocorticoid excess originating from the adrenal or pituitarytumors) or receiving glucocorticoid therapy develop reversible visceralfat obesity. Interestingly, the phenotype of Cushing's syndrome patientsclosely resembles that of Reaven's metabolic syndrome (also known asSyndrome X or insulin resistance syndrome) the symptoms of which includevisceral obesity, glucose intolerance, insulin resistance, hypertension,type 2 diabetes and hyperlipidemia (Reaven (1993) Ann. Rev. Med. 44:121-131). However, the role of glucocorticoids in prevalent forms ofhuman obesity has remained obscure because circulating glucocorticoidconcentrations are not elevated in the majority of metabolic syndromepatients. In fact, glucocorticoid action on target tissue depends notonly on circulating levels but also on intracellular concentration,locally enhanced action of glucocorticoids in adipose tissue andskeletal muscle has been demonstrated in metabolic syndrome. Evidencehas accumulated that enzyme activity of 11βHSD1, which regeneratesactive glucocorticoids from inactive forms and plays a central role inregulating intracellular glucocorticoid concentration, is commonlyelevated in fat depots from obese individuals. This suggests a role forlocal glucocorticoid reactivation in obesity and metabolic syndrome.

Given the ability of 11βHSD1 to regenerate cortisol from inertcirculating cortisone, considerable attention has been given to its rolein the amplification of glucocorticoid function. 11βHSD1 is expressed inmany key GR-rich tissues, including tissues of considerable metabolicimportance such as liver, adipose, and skeletal muscle, and, as such,has been postulated to aid in the tissue-specific potentiation ofglucocorticoid-mediated antagonism of insulin function. Considering a)the phenotypic similarity between glucocorticoid excess (Cushing'ssyndrome) and the metabolic syndrome with normal circulatingglucocorticoids in the latter, as well as b) the ability of 11βHSD1 togenerate active cortisol from inactive cortisone in a tissue-specificmanner, it has been suggested that central obesity and the associatedmetabolic complications in syndrome X result from increased activity of11βHSD1 within adipose tissue, resulting in ‘Cushing's disease of theomentum’ (Bujalska et al. (1997) Lancet 349: 1210-1213). Indeed, 11βHSD1has been shown to be upregulated in adipose tissue of obese rodents andhumans (Livingstone et al. (2000) Endocrinology 131: 560-563; Rask etal. (2001) J. Clin. Endocrinol. Metab. 86: 1418-1421; Lindsay et al.(2003) J. Clin. Endocrinol. Metab. 88: 2738-2744; Wake et al. (2003) J.Clin. Endocrinol. Metab. 88: 3983-3988).

Additional support for this notion has come from studies in mousetransgenic models. Adipose-specific overexpression of 11βHSD1 under thecontrol of the aP2 promoter in mouse produces a phenotype remarkablyreminiscent of human metabolic syndrome (Masuzaki et al. (2001) Science294: 2166-2170; Masuzaki et al. (2003) J. Clinical Invest. 112: 83-90).Importantly, this phenotype occurs without an increase in totalcirculating corticosterone, but rather is driven by a local productionof corticosterone within the adipose depots. The increased activity of11βHSD1 in these mice (2-3 fold) is very similar to that observed inhuman obesity (Rask et al. (2001) J. Clin. Endocrinol. Metab. 86:1418-1421). This suggests that local 11βHSD1-mediated conversion ofinert glucocorticoid to active glucocorticoid can have profoundinfluences whole body insulin sensitivity.

Based on this data, it would be predicted that the loss of 11βHSD1 wouldlead to an increase in insulin sensitivity and glucose tolerance due toa tissue-specific deficiency in active glucocorticoid levels. This is,in fact, the case as shown in studies with 11βHSD1-deficient miceproduced by homologous recombination (Kotelevstev et al. (1997) Proc.Natl. Acad. Sci. 94: 14924-14929; Morton et al. (2001) J. Biol. Chem.276: 41293-41300; Morton et al. (2004) Diabetes 53: 931-938). These miceare completely devoid of 11-keto reductase activity, confirming that11βHSD1 encodes the only activity capable of generating activecorticosterone from inert 11-dehydrocorticosterone. 11βHSD1-deficientmice are resistant to diet- and stress-induced hyperglycemia, exhibitattenuated induction of hepatic gluconeogenic enzymes (PEPCK, G6P), showincreased insulin sensitivity within adipose, and have an improved lipidprofile (decreased triglycerides and increased cardio-protective HDL).Additionally, these animals show resistance to high fat diet-inducedobesity. Taken together, these transgenic mouse studies confirm a rolefor local reactivation of glucocorticoids in controlling hepatic andperipheral insulin sensitivity, and suggest that inhibition of 11βHSD1activity may prove beneficial in treating a number ofglucocorticoid-related disorders, including obesity, insulin resistance,hyperglycemia, and hyperlipidemia.

Data in support of this hypothesis has been published. Recently, it wasreported that 11βHSD1 plays a role in the pathogenesis of centralobesity and the appearance of the metabolic syndrome in humans.Increased expression of the 11βHSD1 gene is associated with metabolicabnormalities in obese women and that increased expression of this geneis suspected to contribute to the increased local conversion ofcortisone to cortisol in adipose tissue of obese individuals (Engeli, etal., (2004) Obes. Res. 12: 9-17).

A new class of 11βHSD1 inhibitors, the arylsulfonamidothiazoles, wasshown to improve hepatic insulin sensitivity and reduce blood glucoselevels in hyperglycemic strains of mice (Barf et al. (2002) J. Med.Chem. 45: 3813-3815; Alberts et al. Endocrinology (2003) 144:4755-4762). Furthermore, it was recently reported that selectiveinhibitors of 11βHSD1 can ameliorate severe hyperglycemia in geneticallydiabetic obese mice. Thus, 11βHSD1 is a promising pharmaceutical targetfor the treatment of the Metabolic Syndrome (Masuzaki, et al., (2003)Curr. Drug Targets Immune Endocr. Metabol. Disord. 3: 255-62).

A. Obesity and Metabolic Syndrome

As described above, multiple lines of evidence suggest that inhibitionof 11βHSD1 activity can be effective in combating obesity and/or aspectsof the metabolic syndrome cluster, including glucose intolerance,insulin resistance, hyperglycemia, hypertension, and/or hyperlipidemia.Glucocorticoids are known antagonists of insulin action, and reductionsin local glucocorticoid levels by inhibition of intracellular cortisoneto cortisol conversion should increase hepatic and/or peripheral insulinsensitivity and potentially reduce visceral adiposity. As describedabove, 11βHSD1 knockout mice are resistant to hyperglycemia, exhibitattenuated induction of key hepatic gluconeogenic enzymes, show markedlyincreased insulin sensitivity within adipose, and have an improved lipidprofile. Additionally, these animals show resistance to high fatdiet-induced obesity (Kotelevstev et al. (1997) Proc. Natl. Acad. Sci.94: 14924-14929; Morton et al. (2001) J. Biol. Chem. 276: 41293-41300;Morton et al. (2004) Diabetes 53: 931-938). Thus, inhibition of 11βHSD1is predicted to have multiple beneficial effects in the liver, adipose,and/or skeletal muscle, particularly related to alleviation ofcomponent(s) of the metabolic syndrome and/or obesity.

B. Pancreatic Function

Glucocorticoids are known to inhibit the glucose-stimulated secretion ofinsulin from pancreatic beta-cells (Billaudel and Sutter (1979) Horm.Metab. Res. 11: 555-560). In both Cushing's syndrome and diabetic Zuckerfa/fa rats, glucose-stimulated insulin secretion is markedly reduced(Ogawa et al. (1992) J. Clin. Invest. 90: 497-504). 11βHSD1 mRNA andactivity has been reported in the pancreatic islet cells of ob/ob miceand inhibition of this activity with carbenoxolone, an 11βHSD1inhibitor, improves glucose-stimulated insulin release (Davani et al.(2000) J. Biol. Chem. 275: 34841-34844). Thus, inhibition of 11βHSD1 ispredicted to have beneficial effects on the pancreas, including theenhancement of glucose-stimulated insulin release.

C. Cognition and Dementia

Mild cognitive impairment is a common feature of aging that may beultimately related to the progression of dementia. In both aged animalsand humans, inter-individual differences in general cognitive functionhave been linked to variability in the long-term exposure toglucocorticoids (Lupien et al. (1998) Nat. Neurosci. 1: 69-73). Further,dysregulation of the HPA axis resulting in chronic exposure toglucocorticoid excess in certain brain subregions has been proposed tocontribute to the decline of cognitive function (McEwen and Sapolsky(1995) Curr. Opin. Neurobiol. 5: 205-216). 11βHSD1 is abundant in thebrain, and is expressed in multiple subregions including thehippocampus, frontal cortex, and cerebellum (Sandeep et al. (2004) Proc.Natl. Acad. Sci. Early Edition: 1-6). Treatment of primary hippocampalcells with the 11βHSD1 inhibitor carbenoxolone protects the cells fromglucocorticoid-mediated exacerbation of excitatory amino acidneurotoxicity (Rajan et al. (1996) J. Neurosci. 16: 65-70).Additionally, 11βHSD1-deficient mice are protected fromglucocorticoid-associated hippocampal dysfunction that is associatedwith aging (Yau et al. (2001) Proc. Natl. Acad. Sci. 98: 4716-4721). Intwo randomized, double-blind, placebo-controlled crossover studies,administration of carbenoxolone improved verbal fluency and verbalmemory (Sandeep et al. (2004) Proc. Natl. Acad. Sci. Early Edition:1-6). Thus, inhibition of 11βHSD1 is predicted to reduce exposure toglucocorticoids in the brain and protect against deleteriousglucocorticoid effects on neuronal function, including cognitiveimpairment, dementia, and/or depression.

D. Intra-Ocular Pressure

Glucocorticoids can be used topically and systemically for a wide rangeof conditions in clinical ophthalmology. One particular complicationwith these treatment regimens is corticosteroid-induced glaucoma. Thispathology is characterized by a significant increase in intra-ocularpressure (IOP). In its most advanced and untreated form, IOP can lead topartial visual field loss and eventually blindness. IOP is produced bythe relationship between aqueous humour production and drainage. Aqueoushumour production occurs in the non-pigmented epithelial cells (NPE) andits drainage is through the cells of the trabecular meshwork. 11βHSD1has been localized to NPE cells (Stokes et al. (2000) Invest.Ophthalmol. Vis. Sci. 41: 1629-1683; Rauz et al. (2001) Invest.Ophthalmol. Vis. Sci. 42: 2037-2042) and its function is likely relevantto the amplification of glucocorticoid activity within these cells. Thisnotion has been confirmed by the observation that free cortisolconcentration greatly exceeds that of cortisone in the aqueous humour(14:1 ratio). The functional significance of 11βHSD1 in the eye has beenevaluated using the inhibitor carbenoxolone in healthy volunteers (Rauzet al. (2001) Invest. Ophthalmol. Vis. Sci. 42: 2037-2042). After sevendays of carbenoxolone treatment, IOP was reduced by 18%. Thus,inhibition of 11βHSD1 in the eye is predicted to reduce localglucocorticoid concentrations and IOP, producing beneficial effects inthe management of glaucoma and other visual disorders.

E. Hypertension

Adipocyte-derived hypertensive substances such as leptin andangiotensinogen have been proposed to be involved in the pathogenesis ofobesity-related hypertension (Matsuzawa et al. (1999) Ann. N.Y. Acad.Sci. 892: 146-154; Wajchenberg (2000) Endocr. Rev. 21: 697-738). Leptin,which is secreted in excess in aP2-11βHSD1 transgenic mice (Masuzaki etal. (2003) J. Clinical Invest. 112: 83-90), can activate varioussympathetic nervous system pathways, including those that regulate bloodpressure (Matsuzawa et al. (1999) Ann. N.Y. Acad. Sci. 892: 146-154).Additionally, the renin-angiotensin system (RAS) has been shown to be amajor determinant of blood pressure (Walker et al. (1979) Hypertension1: 287-291). Angiotensinogen, which is produced in liver and adiposetissue, is the key substrate for renin and drives RAS activation. Plasmaangiotensinogen levels are markedly elevated in aP2-11βHSD1 transgenicmice, as are angiotensin II and aldosterone (Masuzaki et al. (2003) J.Clinical Invest. 112: 83-90). These forces likely drive the elevatedblood pressure observed in aP2-11βHSD1 transgenic mice. Treatment ofthese mice with low doses of an angiotensin II receptor antagonistabolishes this hypertension (Masuzaki et al. (2003) J. Clinical Invest.112: 83-90). This data illustrates the importance of localglucocorticoid reactivation in adipose tissue and liver, and suggeststhat hypertension may be caused or exacerbated by 11βHSD1 activity.Thus, inhibition of 11βHSD1 and reduction in adipose and/or hepaticglucocorticoid levels is predicted to have beneficial effects onhypertension and hypertension-related cardiovascular disorders.

F. Bone Disease

Glucocorticoids can have adverse effects on skeletal tissues. Continuedexposure to even moderate glucocorticoid doses can result inosteoporosis (Cannalis (1996) J. Clin. Endocrinol. Metab. 81: 3441-3447)and increased risk for fractures. Experiments in vitro confirm thedeleterious effects of glucocorticoids on both bone-resorbing cells(also known as osteoclasts) and bone forming cells (osteoblasts).11βHSD1 has been shown to be present in cultures of human primaryosteoblasts as well as cells from adult bone, likely a mixture ofosteoclasts and osteoblasts (Cooper et al. (2000) Bone 27: 375-381), andthe 11βHSD1 inhibitor carbenoxolone has been shown to attenuate thenegative effects of glucocorticoids on bone nodule formation (Bellows etal. (1998) Bone 23: 119-125). Thus, inhibition of 11βHSD1 is predictedto decrease the local glucocorticoid concentration within osteoblastsand osteoclasts, producing beneficial effects in various forms of bonedisease, including osteoporosis.

Small molecule inhibitors of 11βHSD1 are currently being developed totreat or prevent 11βHSD1-related diseases such as those described above.For example, certain amide-based inhibitors are reported in WO2004/089470, WO 2004/089896, WO 2004/056745, and WO 2004/065351.Additional small molecule inhibitors of 11βHSD1 are reported in US2005/0282858, US 2006/0009471, US 2005/0288338, US 2006/0009491, US2006/0004049, US 2005/0288317, US 2005/0288329, US 2006/0122197, US2006/0116382, and US 2006/0122210.

11) INCY0035 (US 2007/0066584)

Antagonists of 11βHSD1 have been evaluated in human clinical trials(Kurukulasuriya, et al., (2003) Curr. Med. Chem. 10: 123-53).

In light of the experimental data indicating a role for 11βHSD1 inglucocorticoid-related disorders, metabolic syndrome, hypertension,obesity, insulin resistance, hyperglycemia, hyperlipidemia, type 2diabetes, androgen excess (hirsutism, menstrual irregularity,hyperandrogenism) and polycystic ovary syndrome (PCOS), therapeuticagents aimed at augmentation or suppression of these metabolic pathways,by modulating glucocorticoid signal transduction at the level of 11βHSD1are desirable.

As evidenced herein, there is a continuing need for new and improveddrugs that target 11βHSD1. The compounds, compositions and methodsdescribed herein help meet this and other needs.

SUMMARY OF THE INVENTION

The present invention provides, inter alia, inhibitors of 11βHSD1 havingFormula I:

or pharmaceutically acceptable salts thereof, wherein the variables aredefined below.

The present invention further provides compositions comprising acompound of Formula I, or a pharmaceutically acceptable salt thereof,and at least one pharmaceutically acceptable carrier.

The present invention further provides methods of inhibiting 11βHSD1 bycontacting the 11βHSD1 with a compound of Formula I, or apharmaceutically acceptable salt thereof.

The present invention further provides methods of inhibiting activity of11βHSD1 comprising contacting the 11βHSD1 with a compound of Formula I,or a pharmaceutically acceptable salt thereof.

The present invention further provides methods of inhibiting theconversion of cortisone to cortisol in a cell comprising contacting thecell with a compound of Formula I, or a pharmaceutically acceptable saltthereof.

The present invention further provides methods of inhibiting theproduction of cortisol in a cell comprising contacting the cell with acompound of Formula I, or a pharmaceutically acceptable salt thereof.

The present invention further provides methods of treating variousdiseases including any one of the following disorders, or anycombination of two or more of the following disorders: obesity;diabetes; glucose intolerance; insulin resistance; hyperglycemia;hypertension; hyperlipidemia; cognitive impairment; depression;dementia; glaucoma; cardiovascular disorders; osteoporosis;inflammation; metabolic syndrome; androgen excess; or polycystic ovarysyndrome (PCOS) in a patient comprising administering to the patient atherapeutically effective amount of a compound of Formula I, or apharmaceutically acceptable salt thereof.

DETAILED DESCRIPTION

The present invention provides, inter alia, inhibitors of 11βHSD1 havingFormula I:

or pharmaceutically acceptable salts thereof, wherein:

R¹ is F, Cl, Br, or I; and

R² and R³ are independently selected from H, C₁₋₆ alkyl, and C₃₋₆cycloalkyl.

In some embodiments:

-   -   R¹ is F and Cl; and    -   R² and R³ are independently selected from H and C₁₋₄ alkyl.

In some embodiments, R¹ is F or Cl.

In some embodiments, R¹ is F.

In some embodiments, R¹ is Cl.

In some embodiments, R² and R³ are independently selected from H,methyl, and ethyl.

In some embodiments, at least one of R² and R³ is other than H.

In some embodiments, the compounds of the invention have Formula II:

At various places in the present specification, substituents ofcompounds of the invention are disclosed in groups or in ranges. It isspecifically intended that the invention include each and everyindividual subcombination of the members of such groups and ranges. Forexample, the term “C₁₋₆ alkyl” is specifically intended to individuallydisclose methyl, ethyl, C₃ alkyl, C₄ alkyl, C₅ alkyl, and C₆ alkyl.

It is further appreciated that certain features of the invention, whichare, for clarity, described in the context of separate embodiments, canalso be provided in combination in a single embodiment. Conversely,various features of the invention which are, for brevity, described inthe context of a single embodiment, can also be provided separately orin any suitable subcombination.

As used herein, the term “alkyl” is meant to refer to a saturatedhydrocarbon group which is straight-chained or branched. Example alkylgroups include methyl (Me), ethyl (Et), propyl (e.g., n-propyl andisopropyl), butyl (e.g., n-butyl, isobutyl, t-butyl), pentyl (e.g.,n-pentyl, isopentyl, neopentyl), and the like.

As used herein, “cycloalkyl” refers to non-aromatic 3-7 memberedcarbocycles including, for example, cyclopropyl, cyclobutyl,cyclopentyl, and cyclohexyl.

The compounds described herein are asymmetric (e.g., having one or morestereocenters). All stereoisomers, such as enantiomers, are intendedunless otherwise indicated. Compounds of the present invention thatcontain asymmetrically substituted carbon atoms can be isolated inoptically active or racemic forms. Methods on how to prepare opticallyactive forms from optically active starting materials are known in theart, such as by resolution of racemic mixtures or by stereoselectivesynthesis. Cis and trans isomers of the compounds of the presentinvention are described and may be isolated as a mixture of isomers oras separated isomeric forms.

Compounds of the invention can also include tautomeric forms. Tautomericforms result from the swapping of a single bond with an adjacent doublebond together with the concomitant migration of a proton. Tautomericforms include prototropic tautomers which are isomeric protonationstates having the same empirical formula and total charge. Exampleprototropic tautomers include ketone—enol pairs, amide—imidic acidpairs, lactam—lactim pairs, amide—imidic acid pairs, enamine—iminepairs, and annular forms where a proton can occupy two or more positionsof a heterocyclic system, for example, 1H- and 3H-imidazole, 1H-, 2H-and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole.Tautomeric forms can be in equilibrium or sterically locked into oneform by appropriate substitution.

Compounds of the invention can also include all isotopes of atomsoccurring in the intermediates or final compounds. Isotopes includethose atoms having the same atomic number but different mass numbers.For example, isotopes of hydrogen include tritium and deuterium.

All compounds, and pharmaceutically acceptable salts thereof, may beobtained in various solid forms, including solvated or hydrated forms.In some embodiments, the solid form is a crystalline form. Methods forpreparing and discovering different solid forms are routine in the artand include, for example, X-ray powder diffraction, differentialscanning calorimetry, thermogravimetric analysis, dynamic vaporsorption, FT-IR, Raman scattering methods, solid state NMR, Karl-Fischertitration, etc.

In some embodiments, the compounds of the invention, and salts thereof,are substantially isolated. By “substantially isolated” is meant thatthe compound is at least partially or substantially separated from theenvironment in which it was formed or detected. Partial separation caninclude, for example, a composition enriched in the compound of theinvention. Substantial separation can include compositions containing atleast about 50%, at least about 60%, at least about 70%, at least about80%, at least about 90%, at least about 95%, at least about 97%, or atleast about 99% by weight of the compound of the invention, or saltthereof. Methods for isolating compounds and their salts are routine inthe art.

The present invention also includes pharmaceutically acceptable salts ofthe compounds described herein. As used herein, “pharmaceuticallyacceptable salts” refers to derivatives of the disclosed compoundswherein the parent compound is modified by converting an existing acidor base moiety to its salt form. Examples of pharmaceutically acceptablesalts include, but are not limited to, mineral or organic acid salts ofbasic residues such as amines; alkali or organic salts of acidicresidues such as carboxylic acids; and the like. The pharmaceuticallyacceptable salts of the present invention include the conventionalnon-toxic salts of the parent compound formed, for example, fromnon-toxic inorganic or organic acids. The pharmaceutically acceptablesalts of the present invention can be synthesized from the parentcompound which contains a basic or acidic moiety by conventionalchemical methods. Generally, such salts can be prepared by reacting thefree acid or base forms of these compounds with a stoichiometric amountof the appropriate base or acid in water or in an organic solvent, or ina mixture of the two; generally, nonaqueous media like ether, ethylacetate, ethanol, isopropanol, or acetonitrile are preferred. Lists ofsuitable salts are found in Remington's Pharmaceutical Sciences, 17thed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal ofPharmaceutical Science, 66, 2 (1977), each of which is incorporatedherein by reference in its entirety.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

Compounds of the invention can modulate activity of 11βHSD1. The term“modulate” is meant to refer to an ability to increase or decreaseactivity of an enzyme or receptor. Accordingly, compounds of theinvention can be used in methods of modulating 11βHSD1 by contacting theenzyme or receptor with any one or more of the compounds or compositionsdescribed herein. In some embodiments, compounds of the presentinvention can act as inhibitors of 11βHSD1. In further embodiments, thecompounds of the invention can be used to modulate activity of 11βHSD1in an individual in need of modulation of the enzyme or receptor byadministering a modulating amount of a compound of the invention.

The present invention further provides methods of inhibiting theconversion of cortisone to cortisol in a cell, or inhibiting theproduction of cortisol in a cell, where conversion to or production ofcortisol is mediated, at least in part, by 11βHSD1 activity. Methods ofmeasuring conversion rates of cortisone to cortisol and vice versa, aswell as methods for measuring levels of cortisone and cortisol in cells,are routine in the art.

The present invention further provides methods of increasing insulinsensitivity of a cell by contacting the cell with a compound of theinvention. Methods of measuring insulin sensitivity are routine in theart.

The present invention further provides methods of treating diseaseassociated with activity or expression, including abnormal activity andoverexpression, of 11βHSD1 in an individual (e.g., patient) byadministering to the individual in need of such treatment atherapeutically effective amount or dose of a compound of the presentinvention, or pharmaceutically acceptable salt thereof, or apharmaceutical composition thereof. Example diseases can include anydisease, disorder or condition that is directly or indirectly linked toexpression or activity of the enzyme. An 11βHSD1-associated disease canalso include any disease, disorder or condition that can be prevented,ameliorated, or cured by modulating enzyme activity.

Examples of 11βHSD1-associated diseases include obesity, diabetes,glucose intolerance, insulin resistance, hyperglycemia, hypertension,hyperlipidemia, cognitive impairment, dementia, depression (e.g.,psychotic depression), glaucoma, cardiovascular disorders, osteoporosis,and inflammation. Further examples of 11βHSD1-associated diseasesinclude metabolic syndrome, type 2 diabetes, androgen excess (hirsutism,menstrual irregularity, hyperandrogenism) and polycystic ovary syndrome(PCOS).

As used herein, the term “cell” is meant to refer to a cell that is invitro, ex vivo or in vivo. In some embodiments, an ex vivo cell can bepart of a tissue sample excised from an organism such as a mammal. Insome embodiments, an in vitro cell can be a cell in a cell culture. Insome embodiments, an in vivo cell is a cell living in an organism suchas a mammal. In some embodiments, the cell is an adipocyte, a pancreaticcell, a hepatocyte, neuron, or cell comprising the eye (ocular cell).

As used herein, the term “contacting” refers to the bringing together ofindicated moieties in an in vitro system or an in vivo system. Forexample, “contacting” the 11βHSD1 enzyme with a compound of theinvention includes the administration of a compound of the presentinvention to an individual or patient, such as a human, having 11βHSD1,as well as, for example, introducing a compound of the invention into asample containing a cellular or purified preparation containing the11βHSD1 enzyme.

As used herein, the term “individual” or “patient,” usedinterchangeably, refers to any animal, including mammals, preferablymice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep,horses, or primates, and most preferably humans.

As used herein, the term “treating” or “treatment” refers to one or moreof (1) preventing the disease; for example, preventing a disease,condition or disorder in an individual who may be predisposed to thedisease, condition or disorder but does not yet experience or displaythe pathology or symptomatology of the disease; (2) inhibiting thedisease; for example, inhibiting a disease, condition or disorder in anindividual who is experiencing or displaying the pathology orsymptomatology of the disease, condition or disorder; and (3)ameliorating the disease; for example, ameliorating a disease, conditionor disorder in an individual who is experiencing or displaying thepathology or symptomatology of the disease, condition or disorder (i.e.,reversing the pathology and/or symptomatology) such as decreasing theseverity of disease.

When employed as pharmaceuticals, the compounds of the invention can beadministered in the form of pharmaceutical compositions which is acombination of a compound of the invention and at least onepharmaceutically acceptable carrier. These compositions can be preparedin a manner well known in the pharmaceutical art, and can beadministered by a variety of routes, depending upon whether local orsystemic treatment is desired and upon the area to be treated.Administration may be topical (including ophthalmic and to mucousmembranes including intranasal, vaginal and rectal delivery), pulmonary(e.g., by inhalation or insufflation of powders or aerosols, includingby nebulizer; intratracheal, intranasal, epidermal and transdermal),ocular, oral or parenteral. Methods for ocular delivery can includetopical administration (eye drops), subconjunctival, periocular orintravitreal injection or introduction by balloon catheter or ophthalmicinserts surgically placed in the conjunctival sac. Parenteraladministration includes intravenous, intraarterial, subcutaneous,intraperitoneal or intramuscular injection or infusion; or intracranial,e.g., intrathecal or intraventricular, administration. Parenteraladministration can be in the form of a single bolus dose, or may be, forexample, by a continuous perfusion pump. Pharmaceutical compositions andformulations for topical administration may include transdermal patches,ointments, lotions, creams, gels, drops, suppositories, sprays, liquidsand powders. Conventional pharmaceutical carriers, aqueous, powder oroily bases, thickeners and the like may be necessary or desirable.

This invention also includes pharmaceutical compositions which contain,as the active ingredient, one or more of the compounds of the inventionabove in combination with one or more pharmaceutically acceptablecarriers. In making the compositions of the invention, the activeingredient is typically mixed with an excipient, diluted by an excipientor enclosed within such a carrier in the form of, for example, acapsule, sachet, paper, or other container. When the excipient serves asa diluent, it can be a solid, semi-solid, or liquid material, which actsas a vehicle, carrier or medium for the active ingredient. Thus, thecompositions can be in the form of tablets, pills, powders, lozenges,sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups,aerosols (as a solid or in a liquid medium), ointments containing, forexample, up to 10% by weight of the active compound, soft and hardgelatin capsules, suppositories, sterile injectable solutions, andsterile packaged powders.

In preparing a formulation, the active compound can be milled to providethe appropriate particle size prior to combining with the otheringredients. If the active compound is substantially insoluble, it canbe milled to a particle size of less than 200 mesh. If the activecompound is substantially water soluble, the particle size can beadjusted by milling to provide a substantially uniform distribution inthe formulation, e.g. about 40 mesh.

The compounds of the invention may be milled using known millingprocedures such as wet milling to obtain a particle size appropriate fortablet formation and for other formulation types. Finely divided(nanoparticulate) preparations of the compounds of the invention can beprepared by processes known in the art, for example see InternationalPatent Application No. WO 2002/000196.

Some examples of suitable excipients include lactose, dextrose, sucrose,sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates,tragacanth, gelatin, calcium silicate, microcrystalline cellulose,polyvinylpyrrolidone, cellulose, water, syrup, and methyl cellulose. Theformulations can additionally include: lubricating agents such as talc,magnesium stearate, and mineral oil; wetting agents; emulsifying andsuspending agents; preserving agents such as methyl- andpropylhydroxy-benzoates; sweetening agents; and flavoring agents. Thecompositions of the invention can be formulated so as to provide quick,sustained or delayed release of the active ingredient afteradministration to the patient by employing procedures known in the art.

The compositions can be formulated in a unit dosage form, each dosagecontaining from about 5 to about 100 mg, more usually about 10 to about30 mg, of the active ingredient. The term “unit dosage forms” refers tophysically discrete units suitable as unitary dosages for human subjectsand other mammals, each unit containing a predetermined quantity ofactive material calculated to produce the desired therapeutic effect, inassociation with a suitable pharmaceutical excipient.

The active compound can be effective over a wide dosage range and isgenerally administered in a pharmaceutically effective amount. It willbe understood, however, that the amount of the compound actuallyadministered will usually be determined by a physician, according to therelevant circumstances, including the condition to be treated, thechosen route of administration, the actual compound administered, theage, weight, and response of the individual patient, the severity of thepatient's symptoms, and the like.

For preparing solid compositions such as tablets, the principal activeingredient is mixed with a pharmaceutical excipient to form a solidpreformulation composition containing a homogeneous mixture of acompound of the present invention. When referring to thesepreformulation compositions as homogeneous, the active ingredient istypically dispersed evenly throughout the composition so that thecomposition can be readily subdivided into equally effective unit dosageforms such as tablets, pills and capsules. This solid preformulation isthen subdivided into unit dosage forms of the type described abovecontaining from, for example, 0.1 to about 500 mg of the activeingredient of the present invention.

The tablets or pills of the present invention can be coated or otherwisecompounded to provide a dosage form affording the advantage of prolongedaction. For example, the tablet or pill can comprise an inner dosage andan outer dosage component, the latter being in the form of an envelopeover the former. The two components can be separated by an enteric layerwhich serves to resist disintegration in the stomach and permit theinner component to pass intact into the duodenum or to be delayed inrelease. A variety of materials can be used for such enteric layers orcoatings, such materials including a number of polymeric acids andmixtures of polymeric acids with such materials as shellac, cetylalcohol, and cellulose acetate.

The liquid forms in which the compounds and compositions of the presentinvention can be incorporated for administration orally or by injectioninclude aqueous solutions, suitably flavored syrups, aqueous or oilsuspensions, and flavored emulsions with edible oils such as cottonseedoil, sesame oil, coconut oil, or peanut oil, as well as elixirs andsimilar pharmaceutical vehicles.

Compositions for inhalation or insufflation include solutions andsuspensions in pharmaceutically acceptable, aqueous or organic solvents,or mixtures thereof, and powders. The liquid or solid compositions maycontain suitable pharmaceutically acceptable excipients as describedsupra. In some embodiments, the compositions are administered by theoral or nasal respiratory route for local or systemic effect.Compositions in can be nebulized by use of inert gases. Nebulizedsolutions may be breathed directly from the nebulizing device or thenebulizing device can be attached to a face masks tent, or intermittentpositive pressure breathing machine. Solution, suspension, or powdercompositions can be administered orally or nasally from devices whichdeliver the formulation in an appropriate manner.

The amount of compound or composition administered to a patient willvary depending upon what is being administered, the purpose of theadministration, such as prophylaxis or therapy, the state of thepatient, the manner of administration, and the like. In therapeuticapplications, compositions can be administered to a patient alreadysuffering from a disease in an amount sufficient to cure or at leastpartially arrest the symptoms of the disease and its complications.Effective doses will depend on the disease condition being treated aswell as by the judgment of the attending clinician depending uponfactors such as the severity of the disease, the age, weight and generalcondition of the patient, and the like.

The compositions administered to a patient can be in the form ofpharmaceutical compositions described above. These compositions can besterilized by conventional sterilization techniques, or may be sterilefiltered. Aqueous solutions can be packaged for use as is, orlyophilized, the lyophilized preparation being combined with a sterileaqueous carrier prior to administration. The pH of the compoundpreparations typically will be between 3 and 11, more preferably from 5to 9 and most preferably from 7 to 8. It will be understood that use ofcertain of the foregoing excipients, carriers, or stabilizers willresult in the formation of pharmaceutical salts.

The therapeutic dosage of the compounds of the present invention canvary according to, for example, the particular use for which thetreatment is made, the manner of administration of the compound, thehealth and condition of the patient, and the judgment of the prescribingphysician. The proportion or concentration of a compound of theinvention in a pharmaceutical composition can vary depending upon anumber of factors including dosage, chemical characteristics (e.g.,hydrophobicity), and the route of administration. For example, thecompounds of the invention can be provided in an aqueous physiologicalbuffer solution containing about 0.1 to about 10% w/v of the compoundfor parenteral administration. Some typical dose ranges are from about 1μg/kg to about 1 g/kg of body weight per day. In some embodiments, thedose range is from about 0.01 mg/kg to about 100 mg/kg of body weightper day. The dosage is likely to depend on such variables as the typeand extent of progression of the disease or disorder, the overall healthstatus of the particular patient, the relative biological efficacy ofthe compound selected, formulation of the excipient, and its route ofadministration. Effective doses can be extrapolated from dose-responsecurves derived from in vitro or animal model test systems.

The compounds of the invention can also be formulated in combinationwith one or more additional active ingredients which can include anypharmaceutical agent such as anti-viral agents, vaccines, antibodies,immune enhancers, immune suppressants, anti-inflammatory agents,analgesics, and drugs for the treatment of diabetes or obesity,hyperglycemia, hypertension, hyperlipidemia, and the like. Agents fortreatment of metabolic disorders with which a compound of the inventioncould be combined include, but are not limited to, amylin analogues,incretin mimetics, inhibitors of the incretin-degrading enzymedipeptidyl peptidase-IV, agonists of peroxisome proliferator-activatedreceptor (PPAR)-a and PPAR-g, and CB1 cannabinoid receptor inhibitors.

The invention will be described in greater detail by way of specificexamples. The following examples are offered for illustrative purposes,and are not intended to limit the invention in any manner. Those ofskill in the art will readily recognize a variety of noncriticalparameters which can be changed or modified to yield essentially thesame results.

EXAMPLES

All compounds were purified by either flash column chromatography orreversed-phase liquid chromatography using a Waters FractionLynx LC-MSsystem with mass directed fractionation. Column: Waters XBridge C₁₈ 5μm, 19×100 mm; mobile phase A: 0.15% NH₄OH in water and mobile phase B:0.15% NH₄OH in acetonitrile; the flow rate was 30 ml/m, the separatinggradient was optimized for each compound using the Compound SpecificMethod Optimization protocol as described in literature [“PreparativeLC-MS Purification: Improved Compound Specific Method Optimization”, KBlom, B. Glass, R. Sparks, A. Combs, J. Combi. Chem., 2004, 6, 874-883].

The separated product was then typically subjected to analytical LC/MSfor purity check under the following conditions: Instrument; Agilent1100 series, LC/MSD, Column: Waters Sunfire™ C₁₈ 5 μm, 2.1×5.0 mm,Buffers: mobile phase A: 0.025% TFA in water and mobile phase B: 0.025%TFA in acetonitrile; gradient 2% to 80% of buffer B in 3 min with flowrate 1.5 mL/min.

Example 15-{3-Fluoro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}-N-methylpyridine-2-carboxamide

Step 1: 1-benzyl 3-ethyl piperidine-1,3-dicarboxylate

Benzyl chloroformate (Aldrich, cat #:119938) (191 mL, 1.34 mol) wasslowly added to a cooled (at 0° C.) mixture of ethylpiperidine-3-carboxylate (Aldrich, cat #:194360) (200 g, 1.27 mol) andtriethylamine (266 mL, 1.91 mol) in methylene chloride (1000 mL). Thereaction mixture was allowed to gradually warm to ambient temperatureand stirred for 3 h. The reaction was quenched by the addition of 1N HClaq. solution and the product was extracted several times with methylenechloride. The combined extracts were washed with water, saturated aq.NaHCO₃, water, brine, dried over MgSO₄, filtered and concentrated underreduced pressure to afford the desired product as oil (359.8 g, 97%).LC/MS 292.2 (M+H)⁺.

Step 2: 1-benzyl 3-ethyl3-(3-methylbut-2-en-1-yl)piperidine-1,3-dicarboxylate

To a solution of 1-benzyl 3-ethyl piperidine-1,3-dicarboxylate (120.0 g,0.412 mol) in THF (400 ml) cooled at −78° C. was added dropwise 270 mLof sodium bis(trimethylsilyl)amide solution (1M solution in THF fromAldrich, cat#:245585) over 2 h. The mixture was stirred at −78° C. foradditional 1 h. Then 1-bromo-3-methylbut-2-ene (Aldrich cat #: 249904)(71 mL, 0.62 mol) was added slowly over 1 h. The mixture was stirred at−78° C. for 30 min, and allowed to warm to r.t. and stirred for anadditional 3 h. The reaction mixture was quenched with 1N HCl aq.solution. Most of THF was removed under reduced pressure. The residuewas extracted with ethyl acetate. The combined extracts were washed withsat. aq. NaHCO₃ and brine, then dried over MgSO₄, filtered andconcentrated under reduced pressure. The crude residue was purified byflash column chromatography on a silica gel column with 10˜20% ethylacetate in hexane to yield the desired product (140 g, 94%). LC/MS:m/e=332.2 (M+H)⁺.

Step 3: 1-benzyl 3-ethyl 3-(2-oxoethyl)piperidine-1,3-dicarboxylate

Ozone was passed through a solution of 1-benzyl 3-ethyl3-(3-methylbut-2-en-1-yl)piperidine-1,3-dicarboxylate (35.2 g, 0.0979mol) in methylene chloride (800 mL) at −78° C. until the color of thesolution turned blue. The reaction mixture was then flushed withnitrogen until the blue color dissipated. Dimethylsulfide (Aldrich, cat#: 274380) (14 mL, 0.19 mol) and triethylamine (26.5 mL, 0.19 mol) wereadded and the mixture was stirred at ambient temperature overnight. Thevolatile solvent were removed under reduced pressure and purifieddirectly by flash chromatography on a silica gel column with 20% ethylacetate in hexanes to afford the desired product in quantitative yield.LC/MS 334.2 (M+H)⁺.

Step 4: Benzyl2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]decane-7-carboxylate

To a suspension of cis-4-aminocyclohexanol hydrochloride (Available fromSijia Medchem Lab, China) (13.8 g, 0.0910 mol) and 1-benzyl 3-ethyl3-(2-oxoethyl)piperidine-1,3-dicarboxylate (31.0 g, 0.0930 mol) in1,2-dichloroethane (250 mL) was added triethylamine (23.3 mL, 0.167 mol)at room temperature. The mixture was stirred at 40° C. overnight. Sodiumtriacetoxyborohydride (Aldrich, cat #: 316393) (49.3 g, 0.232 mol) wasadded to the above mixture and stirred at r.t. for 1 h. LC/MS dataindicated that the starting material was consumed, and an intermediateproduct with m/e: 433.2 (M+H)⁺ was observed.

The mixture was then heated at 80° C. for 4 h or until LC/MS showed theintermediate amine (m/e: 433.2) was consumed. The reaction mixture wasquenched with aq. NaHCO₃. The organic layer was washed with brine, driedover MgSO₄, filtered and concentrated under reduced pressure. The crudematerial was dried under reduced pressure overnight to give colorlessviscous oil (26.9 g, 66.8%). LC/MS m/e 387.2 (M+H)⁺.

Step 5: Benzyl(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]decane-7-carboxylate

The racemic mixture obtained from above step (26.9 g) was purified on anAgilent 1100 series preparatory system using a Chiralcel OD-H column(3.0×25 cm, 5 micron particle size, Chiral Technologies) eluting with30% ethanol/hexanes (isocratic, 22 mL/min.). The column loading wasapproximately 150 mg/injection and peak collection was triggered by UVabsorbance at 220 nM. Peak 1 eluted at approximately at 8.5 min. andPeak 2 eluted at approximately 9.8 min. The fractions of Peak 2 werecombined and concentrated to provide the desired product (11.9 g) as awhite foamy solid. The optical purity of the pooled material from peak 2was determined by using an Agilent 1100 series analytical systemequipped with a Chiralcel OD-H column (4.6×250 mm, 5 micron particlesize, Chiral Technologies) and eluting with 30% ethanol/hexanes(isocratic, 0.8 mL/min.). LC/MS m/e 387.2 (M+H)⁺. The absolutestereochemistry of the peak 2 was established based on X-ray singlecrystal structure determination of close analogs: Benzyl(5S)-2-(trans-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]decane-7-carboxylateand (5S)-2-(cis-4-{[tert-butyl(dimethyl)silyl]oxy}cyclohexyl)-2,7-diazaspiro[4.5]decan-1-one prepared asdescribed in Steps 5a-c.

Step 5a: Benzyl2-(cis-4-{[tert-butyl(dimethyl)silyl]oxy}cyclohexyl)-1-oxo-2,7-diazaspiro[4.5]decane-7-carboxylate

To a stirred solution of benzyl2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]decane-7-carboxylate(60.00 g, 155.2 mmol) in anhydrous N,N-dimethylformamide (160 mL) atr.t. was added 1H-imidazole (32.0 g, 466 mmol) andtert-butyldimethylsilyl chloride (36.2 g, 233 mmol). The reactionmixture was stirred at r.t. for 4 h, quenched with water (150 mL), andextracted with EtOAc (3×150 mL). The combined organic layers were washedwith brine, dried over sodium sulfate, filtered and concentrated underreduced pressure to afford the crude product (84 g). The pure product(55.4 g) was obtained by re-crystallization of the crude product fromheptane. The mother liquor was concentrated and subjected topurification by flash chromatography on a silical gel column elutingwith AcOEt/Haxane to give additional 14.4 g of the product with a total89.7% yield.

Step 5b:2-(cis-4-{[tert-butyl(dimethyl)silyl]oxy}cyclohexyl)-2,7-diazaspiro[4.5]decan-1-one

To a solution of benzyl2-(cis-4-{[tert-butyl(dimethyl)silyl]oxy}cyclohexyl)-1-oxo-2,7-diazaspiro[4.5]decane-7-carboxylate(18.0 g, 35.9 mmol) in methanol (150 mL) was added 10% palladium oncarbon (Aldrich, cat #: 520888) (1.8 g, 1.5 mmol) under the atmosphereof nitrogen. The reaction mixture was hydrogenated and shaken at 50 psifor 20 h. The reaction mixture was filtered through a pad of Celite andthen washed with methanol (300 mL). The filtrate was concentrated underreduced pressure to give the desired product as a white solid inquantitative yield.

Step 5c:(5S)-2-(cis-4-{[tert-butyl(dimethyl)silyl]oxy}cyclohexyl)-2,7-diazaspiro[4.5]decan-1-one

2-(cis-4-{[tert-Butyl(dimethyl)silyl]oxy}cyclohexyl)-2,7-diazaspiro[4.5]decan-1-one(7.00 g, 19.1 mmol) was dissolved in acetonitrile (50 mL) and methanol(7 mL) at r.t. After the starting material was completely dissolved, thesolution was heated up to 70° C. To the above solution was slowly addeda solution of (2R)-hydroxy(phenyl)acetic acid (1.45 g, 9.55 mmol) inacetonitrile (20 mL) at 65-70° C. After addition, the solution washeated at 74° C. for 10 min, and allowed to cool slowly to roomtemperature overnight. The crystalline formed was collected byfiltration to afford 3.38 g of the desired product as(2R)-hydroxy(phenyl)acetic acid salt. The resulting salt (3.38 g) wasdissolved in water (50 mL), and adjusted to pH-12 with 40 mL aq K₂CO₃solution (2.0 M). The mixture was extracted with dichloromethane (3times). The combined organic layers were dried with magnesium sulfate,filtered, and concentrated under reduced pressure to afford the desiredproduct as a free base (colorless crystalline solid) (2.37 g). Theabsolute stereochemistry of this compound was established by X-raysingle crystal structure determination of (2R)-hydroxy(phenyl)aceticacid salt of(5S)-2-(cis-4-{[tert-butyl(dimethyl)silyl]oxy}cyclohexyl)-2,7-diazaspiro[4.5]decan-1-one.

Step 6: (5S)-2-(cis-4-hydroxycyclohexyl)-2,7-diazaspiro[4.5]decan-1-one

Benzyl(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]decane-7-carboxylateprepared in Step 5 (0.266 g, 0.000688 mol) was dissolved in methanol(5.0 mL) and stirred under an atmosphere of hydrogen in the presence of10% palladium on carbon (Aldrich, cat #: 520888) (20.0 mg) at r.t. for 2h. The reaction mixture was filtered and the volatile solvents wereremoved under reduced pressure to afford the desired product inquantitative yield. LC/MS m/e 253.2 (M+H)⁺.

Step 7:(5S)-7-(4-bromo-2-fluorophenyl)-2-(cis-4-hydroxycyclohexyl)-2,7-diazaspiro[4.5]decan-1-one

A mixture of(5S)-2-(cis-4-hydroxycyclohexyl)-2,7-diazaspiro[4.5]decan-1-one (1.04 g,0.00412 mol), 4-bromo-2-fluoro-1-iodobenzene (Aldrich, cat #: 283304)(1.85 g, 0.00615 mol), copper(I) iodide (Aldrich, cat #: 215554) (0.122g, 0.000640 mol), potassium phosphate (2.63 g, 0.0124 mol) and1,2-ethanediol (0.48 mL, 0.0086 mol) in 1-butanol (3.90 mL) was heatedat 100° C. under nitrogen for 2 d. The reaction was quenched with water,and extracted with ether. The organic layers were combined, washed withwater, brine, dried over Na₂SO₄, and filtered. The filtrate wasevaporated under reduced pressure. The residue was purified by flashcolumn chromatography on a silica gel column eluting with 0 to 5%methanol in DCM to yield the desired product (950 mg, 54.2%). LC/MS m/e425.1/427.0 (M+H)⁺.

Step 8:5-3-fluoro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl-N-methylpyridine-2-carboxamide

Potassium phosphate (637 mg, 0.00300 mol) in water (3.00 mL) was addedto a mixture of(5S)-7-(4-bromo-2-fluorophenyl)-2-(cis-4-hydroxycyclohexyl)-2,7-diazaspiro[4.5]decan-1-one(425 mg, 0.00100 mol),N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carboxamide(Frontier Inc., cat #: M10074) (393 mg, 0.00150 mol) andtetrakis(triphenylphosphine)palladium (Aldrich, cat #: 216666) (35 mg,0.000030 mol) in 1,4-dioxane (3.00 mL). The resulting mixture was heatedat 120° C. for 24 h. The mixture was diluted with ethyl acetate andwashed with water and brine. The organic layer was dried over Na₂SO₄,filtered, concentrated under reduced pressure. The residue was purifiedby flash column chromatography on a silica gel column eluting with 5%methanol in DCM to yield the desired product (285 mg, 59.3%). LC/MS m/e481.2 (M+H)⁺. ¹H-NMR (400 MHz, DMSO-d₆): 8.89 (1H, dd, J=2.5, 0.6 Hz),8.76 (1H, q, J=4.7 Hz), 8.22 (1H, dd, J=8.4, 2.5 Hz), 8.03 (1H, dd,J=8.4, 0.6 Hz), 7.65 (1H, dd, J=14.2, 2.1 Hz), 7.56 (1H, dd, J=8.5, 2.1Hz), 7.13 (1H, t, J=8.5 Hz), 4.37 (1H, d, J=3.1 Hz), 3.78 (1H, m), 3.71(1H, m), 3.21-3.38 (3H, m), 3.07 (1H, d, J=11.4 Hz), 2.81 (3H, d, J=4.7Hz), 2.64-2.74 (2H, m), 2.18-2.26 (1H, m), 1.60-1.91 (8H, m), 1.39-1.51(3H, m), 1.21-1.30 (2H, m).

Example 25-{3-Fluoro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}-N,N-dimethylpyridine-2-carboxamide

Step 1: 5-bromo-N,N-dimethylpyridine-2-carboxamide

Oxalyl chloride (20.0 mL, 0.236 mol) was added to a solution of5-bromopyridine-2-carboxylic acid (Alfa Aesar, cat #: B25675) (10.1 g,0.0500 mol) in methylene chloride (60 mL) at r.t. followed by 5 drops ofDMF. The mixture was stirred at r.t. for 2 h. The volatiles wereevaporated under reduced pressure. The residue was azotropicallyevaporated with toluene twice. The residue was then dissolved in DCM (30mL) followed by the addition of 30 mL of dimethylamine in THF solution(2.0 M) (Aldrich, cat #: 391956) and Hunig's base (20.0 mL) (Aldrich,cat #: 496219). The mixture was stirred at r.t. for 3 h. The reactionmixture was diluted with DCM (100 mL) and washed with water, 1N HCl andbrine. The organic phase was dried over Na₂SO₄, filtered andconcentrated to give the desired product (10.5 g, 91.7%).

Step 2:N,N-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carboxamide

A mixture of 5-bromo-N,N-dimethylpyridine-2-carboxamide (5.73 g, 0.0250mol), 4,4,5,5,4′,4′,5′,5′-octamethyl-[2,2′]bi[[1,3,2]dioxaborolanyl](6.98 g, 0.0275 mol) (Aldrich, cat #: 473294),[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) complexedwith dichloromethane (1:1) (0.6 g, 0.0007 mol) (Aldrich, cat #: 379670),1,1′-bis(diphenylphosphino)ferrocene (0.4 g, 0.8 mmol) (Aldrich, cat #:177261), and potassium acetate (7.36 g, 0.0750 mol) in 1,4-dioxane (100mL) was heated at 120° C. for 20 h. After cooling, the mixture wasconcentrated, diluted with ethyl acetate and washed with sat′d NH₄Clsolution, water, brine; dried over Na₂SO₄. After filtration, thefiltrate was concentrated and the crude material was further purified ona silica gel column eluting with ethyl acetate/hexane to give thedesired product (4.7 g, 68%).

Step 3:5-{3-Fluoro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}-N,N-dimethylpyridine-2-carboxamide

This compound was prepared by using procedures that were analogous tothose described for the synthesis of Example 1, Step 8 starting fromN,N-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carboxamideand(5S)-7-(4-bromo-2-fluorophenyl)-2-(cis-4-hydroxycyclohexyl)-2,7-diazaspiro[4.5]decan-1-one.LC/MS m/e 495.3 (M+H)⁺. ¹H-NMR (400 MHz, DMSO-d₆): 8.86 (1H, d, J=1.7Hz), 8.15 (1H, dd, J=8.1, 2.3 Hz), 7.51-7.65 (3H, m), 7.12 (1H, t, J=8.9Hz), 4.37 (1H, d, J=3.1 Hz), 3.78 (1H, m), 3.71 (1H, m), 3.22-3.38 (3H,m), 3.06 (1H, d, J=11.7 Hz), 3.00 (3H, s), 2.97 (3H, s), 2.64-2.74 (2H,m), 2.18-2.27 (1H, m), 1.60-1.91 (8H, m), 1.39-1.51 (3H, m), 1.22-1.30(2H, m).

Example 3N-Ethyl-5-{3-fluoro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}pyridine-2-carboxamide

Step 1:N-ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carboxamide

This compound was prepared by using procedures that were analogous tothose described for the synthesis of Example 2, Steps 1 & 2 startingfrom 5-bromopyridine-2-carboxylic acid.

Step 2:N-Ethyl-5-{3-fluoro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}pyridine-2-carboxamide

This compound was prepared by using procedures that were analogous tothose described for the synthesis of Example 1, Step 8 starting fromN-ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carboxamideand(5S)-7-(4-bromo-2-fluorophenyl)-2-(cis-4-hydroxycyclohexyl)-2,7-diazaspiro[4.5]decan-1-on.LC/MS m/e 495.3 (M+H)⁺.

Example 45-{3-Chloro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}-N-ethylpyridine-2-carboxamide

Step 1:(5S)-7-(4-bromo-2-chlorophenyl)-2-(cis-4-{[tert-butyl(dimethyl)silyl]oxy}cyclohexyl)-2,7-diazaspiro[4.5]decan-1-one

A mixture of(5S)-2-(cis-4-{[tert-butyl(dimethyl)silyl]oxy}cyclohexyl)-2,7-diazaspiro[4.5]decan-1-one(0.282 g, 0.000769 mol), 4-bromo-2-chloro-1-iodobenzene (0.293 g,0.000922 mol) (Lancaster, cat #: 19245), copper(I) iodide (0.015 g,0.000077 mol), potassium phosphate (0.490 g, 0.00231 mol) and1,2-ethanediol (0.0857 mL, 0.00154 mol) in 1-butanol (0.75 mL) washeated at 100° C. under nitrogen for 2 d. The reaction mixture wasfiltered, concentrated under reduced pressure, and the residue waspurified by flash chromatography on a silica gel column (eluting with 0to 50% ethyl acetate in hexanes) to afford the desired product.

Step 2:5-{3-chloro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}-N-ethylpyridine-2-carboxamide

To a stirred mixture of(5S)-7-(4-bromo-2-chlorophenyl)-2-(cis-4-{[tert-butyl(dimethyl)silyl]oxy}cyclohexyl)-2,7-diazaspiro[4.5]decan-1-one(20 mg, 0.00004 mol),[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) complex withdichloromethane (1:1) (2.0 mg), tetrakis(triphenylphosphine)palladium(1.0 mg) and potassium carbonate (14.9 mg, 0.000108 mol) in anhydrousN,N-dimethylformamide (1 mL) was addedN-ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carboxamide(14.5 mg, 0.054 mmol). The resulting reaction mixture was heated at 150°C. and stirred overnight, followed by the removal of TBS protectinggroup by the addition of 1.7 M of fluorosilicic acid in water (0.10 mL)and the mixture was stirred at r.t. overnight. The reaction mixture wasthen directly purified by RP-HPLC to afford the desired product. LC/MSm/e 511.2 (M+H)⁺. ¹H-NMR (400 MHz, DMSO-d₆): 8.92 (1H, d, J=2.3 Hz),8.84 (1H, t, J=5.9 Hz), 8.26 (1H, dd, J=8.2, 2.3 Hz), 8.06 (1H, d, J=8.2Hz), 7.89 (1H, d, J=2.2 Hz), 7.74 (1H, dd, J=8.5, 2.2 Hz), 7.30 (1H, t,J=8.5 Hz), 4.39 (1H, d, J=3.1 Hz), 3.80 (1H, m), 3.72 (1H, m), 3.24-3.44(5H, m), 3.01 (1H, d, J=11.4 Hz), 2.63-2.74 (2H, m), 2.40-2.53 (1H, m),1.64-1.91 (8H, m), 1.41-1.53 (3H, m), 1.20-1.32 (2H, m), 1.13 (3H, t,J=7.2 Hz).

Example 5 Enzymatic assay of 11βHSD1

All in vitro assays were performed with clarified lysates as the sourceof 11βHSD1 activity. HEK-293 transient transfectants expressing anepitope-tagged version of full-length human 11βHSD1 were harvested bycentrifugation. Roughly 2×10⁷ cells were resuspended in 40 mL of lysisbuffer (25 mM Tris-HCl, pH 7.5, 0.1 M NaCl, 1 mM MgCl₂ and 250 mMsucrose) and lysed in a microfluidizer. Lysates were clarified bycentrifugation and the supernatants were aliquoted and frozen.

Inhibition of 11βHSD1 by test compounds was assessed in vitro by aScintillation Proximity Assay (SPA). Dry test compounds were dissolvedat 5 mM in DMSO. These were diluted in DMSO to suitable concentrationsfor the SPA assay. 0.8 μL of 2-fold serial dilutions of compounds weredotted on 384 well plates in DMSO such that 3 logs of compoundconcentration were covered. 20 μL of clarified lysate was added to eachwell. Reactions were initiated by addition of 20 μL ofsubstrate-cofactor mix in assay buffer (25 mM Tris-HCl, pH 7.5, 0.1 MNaCl, 1 mM MgCl₂) to final concentrations of 400 μM NADPH, 25 nM³H-cortisone and 0.007% Triton X-100. Plates were incubated at 37° C.for one hour. Reactions were quenched by addition of 40 μL of anti-mousecoated SPA beads that had been pre-incubated with 10 μM carbenoxoloneand a cortisol-specific monoclonal antibody. Quenched plates wereincubated for a minimum of 30 minutes at RT prior to reading on aTopcount scintillation counter. Controls with no lysate, inhibitedlysate, and with no mAb were run routinely. Roughly 30% of inputcortisone is reduced by 11βHSD1 in the uninhibited reaction under theseconditions.

Example 6 Cell-Based Assay for 11βHSD1 Activity

Peripheral blood mononuclear cells (PBMCs) were isolated from normalhuman volunteers by Ficoll density centrifugation. Cells were plated at4×10⁵ cells/well in 200 μL of AIM V (Gibco-BRL) media in 96 well plates.The cells were stimulated overnight with 50 ng/ml recombinant human IL-4(R&D Systems). The following morning, 200 nM cortisone (Sigma) was addedin the presence or absence of various concentrations of compound. Thecells were incubated for 48 hours and then supernatants were harvested.Conversion of cortisone to cortisol was determined by a commerciallyavailable ELISA (Assay Design).

Various modifications of the invention, in addition to those describedherein, will be apparent to those skilled in the art from the foregoingdescription. Such modifications are also intended to fall within thescope of the appended claims. Each reference, including all patent,patent applications, and publications, cited in the present applicationis incorporated herein by reference in its entirety.

What is claimed is:
 1. A compound of Formula I:

or a pharmaceutically acceptable salt thereof, wherein: R¹ is F, Cl, Br,or I; and R² and R³ are independently selected from H, C₁₋₆ alkyl, andC₃₋₆ cycloalkyl.
 2. A compound according to claim 1, or apharmaceutically acceptable salt thereof, wherein R¹ is F or Cl.
 3. Acompound according to claim 1, or a pharmaceutically acceptable saltthereof, wherein R² and R³ are independently selected from H and C₁₋₄alkyl.
 4. A compound according to claim 1, or a pharmaceuticallyacceptable salt thereof, wherein at least one of R² and R³ is other thanH.
 5. A compound according to claim 1, or a pharmaceutically acceptablesalt thereof, wherein R¹ is F or Cl, R² and R³ are independentlyselected from H and C₁₋₄ alkyl, and at least one of R² and R³ is otherthan H.
 6. A compound according to claim 1 of Formula II:

or a pharmaceutically acceptable salt thereof.
 7. A compound accordingto claim 6, or a pharmaceutically acceptable salt thereof, wherein R¹ isF or Cl.
 8. A compound according to claim 6, or a pharmaceuticallyacceptable salt thereof, wherein R² and R³ are independently selectedfrom H and C₁₋₄ alkyl.
 9. A compound according to claim 6, or apharmaceutically acceptable salt thereof, wherein at least one of R² andR³ is other than H.
 10. A compound according to claim 6, or apharmaceutically acceptable salt thereof, wherein R¹ is F or Cl, R² andR³ are independently selected from H and C₁₋₄ alkyl, and at least one ofR² and R³ is other than H.
 11. A compound according to claim 1 selectedfrom the following compounds, and pharmaceutically acceptable saltsthereof:5-{3-Fluoro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}-N,N-dimethylpyridine-2-carboxamide;5-{3-Fluoro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}-N-methylpyridine-2-carboxamide;N-Ethyl-5-{3-fluoro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}pyridine-2-carboxamide;and5-{3-Chloro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}-N-ethylpyridine-2-carboxamide.12. A pharmaceutical composition comprising a compound according toclaim 1, or a pharmaceutically acceptable salt thereof, and at least onepharmaceutically acceptable carrier.
 13. A pharmaceutical compositioncomprising a compound according to claim 11, or a pharmaceuticallyacceptable salt thereof, and at least one pharmaceutically acceptablecarrier.
 14. A method of treating obesity; diabetes; glucoseintolerance; insulin resistance; hyperglycemia; hypertension;hyperlipidemia; cognitive impairment; depression; dementia; glaucoma;cardiovascular disorders; osteoporosis; inflammation; metabolicsyndrome; androgen excess; or polycystic ovary syndrome (PCOS) in apatient, comprising administering to said patient a therapeuticallyeffective amount of a compound of claim 1, or a pharmaceuticallyacceptable salt thereof.
 15. A method according to claim 14, wherein thecompound is selected from the following compounds, and pharmaceuticallyacceptable salts thereof:5-{3-Fluoro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}-N,N-dimethylpyridine-2-carboxamide;5-{3-Fluoro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}-N-methylpyridine-2-carboxamide;N-Ethyl-5-{3-fluoro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}pyridine-2-carboxamide;and5-{3-Chloro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}-N-ethylpyridine-2-carboxamide.16. A method according to claim 14, wherein the compound is thefollowing compound, or a pharmaceutically acceptable salt thereof:5-{3-Fluoro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}-N,N-dimethylpyridine-2-carboxamide.17. A method of treating type II diabetes in a patient comprisingadministering to said patient a therapeutically effective amount of acompound of claim 1, or a pharmaceutically acceptable salt thereof. 18.A method according to claim 17, wherein the compound is selected fromthe following compounds, and pharmaceutically acceptable salts thereof:5-{3-Fluoro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}-N,N-dimethylpyridine-2-carboxamide;5-{3-Fluoro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}-N-methylpyridine-2-carboxamide;N-Ethyl-5-{3-fluoro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}pyridine-2-carboxamide;and5-{3-Chloro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}-N-ethylpyridine-2-carboxamide.19. A method according to claim 18, wherein the compound is thefollowing compound, or a pharmaceutically acceptable salt thereof:5-{3-Fluoro-4-[(5S)-2-(cis-4-hydroxycyclohexyl)-1-oxo-2,7-diazaspiro[4.5]dec-7-yl]phenyl}-N,N-dimethylpyridine-2-carboxamide.